Primers and probes for detecting hepatitis c virus

Abstract

The present invention relates to primers, probes, primer sets, primer and probe sets, methods and kits for detecting Hepatitis C virus in a test sample.

1 . A primer for amplifying Hepatitis C virus in a test sample, wherein the primer has a sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, and complements thereof. 
     
     
         2 . A probe for detecting Hepatitis C virus in a test sample, wherein the probe has a sequence selected from the group consisting of: SEQ ID NO:7, SEQ ID NO:8, and complements thereof. 
     
     
         3 . A primer set for amplifying Hepatitis C virus in a test sample, the primer set comprising:
 (a) at least one forward primer having a sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, complements thereof, and any combinations thereof; and   (b) at least one reverse primer having a sequence selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, complements thereof, and any combinations thereof.   
     
     
         4 . A primer and probe set for detecting Hepatitis C virus in a test sample, comprising:
 (a) two forward primers having a sequence of: SEQ ID NO:1 and SEQ ID NO:2, or complements thereof, and four reverse primers having a sequence of: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, or complements thereof; and   (b) two probes having a sequence of: SEQ ID NO:7 and SEQ ID NO:8, or complements thereof.   
     
     
         5 . A method for detecting Hepatitis C virus in a test sample, the method comprising the steps of:
 (a) contacting a test sample with at least one forward primer having a sequence selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:2, or complements thereof and at least one reverse primer having a sequence selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 or complements thereof under amplification conditions to generate a first target sequence; and   (b) detecting hybridization between the first target sequence and at least one probe as an indication of the presence of Hepatitis C virus in the test sample, wherein the at least one probe has a sequence selected from the group consisting of: SEQ ID NO:7 or SEQ ID NO:8, or complements thereof.   
     
     
         6 . The method of  claim 5 , wherein the amplification conditions comprise submitting the test sample to an amplification reaction carried out in the presence of suitable amplification reagents. 
     
     
         7 . The method of  claim 6 , wherein the amplification reaction comprises at least one of:
 a) PCR;   b) real-time PCR; or   c) reverse-Transcriptase PCR (RT-PCR).   
     
     
         8 . The method of  claim 5 , wherein, the at least one probe is labeled with a detectable label. 
     
     
         9 . The method of  claim 8 , wherein the detectable label is directly attached to the at least one probe. 
     
     
         10 . The method of  claim 8 , wherein the detectable label is indirectly attached to the at least one probe. 
     
     
         11 . The method of  claim 8 , wherein the detectable label is directly detectable. 
     
     
         12 . The method of  claim 8 , wherein the detectable label is indirectly detectable. 
     
     
         13 . The method of  claim 8 , wherein the detectable label comprises a fluorescent moiety attached at a 5′ end of the at least one probe. 
     
     
         14 . The method of  claim 13 , wherein the at least one probe further comprises a quencher moiety attached at a 3′ end of the at least one probe. 
     
     
         15 . The method of  claim 5 , further comprising the steps of:
 (a) contacting the test sample with a forward primer having a sequence of SEQ ID NO:1 or a complement thereof and a reverse primer having a sequence of SEQ ID NO:3 or a complement thereof under amplification conditions to generate a second target sequence; and   (b) detecting hybridization between the second target sequence and a probe having a sequence of SEQ ID NO:7 or a complement thereof as an indication of the presence of HCV in the test sample.   
     
     
         16 . A kit for detecting Hepatitis C virus in a test sample, the kit comprising:
 (a) at least one forward primer having a sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, complements thereof, and any combinations thereof;   (b) at least one reverse primer having a sequence selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, complements thereof, and any combinations thereof; and   (c) amplification reagents.   
     
     
         17 . The kit of  claim 16 , further comprising at least one probe, wherein the at least one probe is selected from the group consisting of: SEQ ID NO:7, SEQ ID NO:8, and complements thereof.